Fig. 2
Illustrated potency assay strategy. Ten randomly obtained buffy coats from healthy donors can be processed in parallel to isolate approximately 1 × 109 peripheral blood mononuclear cells (PBMCs) per donor, pooled, labeled with carboxyfluorescein (CFSE) and cryopreserved in appropriate aliquots (e.g., 200 aliquots of 1 × 107 pre-labeled pooled PBMCs (pPBMC)) for subsequent off-the-shelf use as responder cells in the potency assay. Individual mesenchymal stem/progenitor cells (MSPC) from donor or organ origin of choice (e.g., bone marrow (BM), white adipose tissue (WAT), umbilical cord (UC); color code corresponding to Fig. 1) can be tested off-the-shelf or after rescue culture (with or without gamma irradiation (±Rx)) to test their potency to inhibit mitogen (e.g., PHA)-driven pPBMC proliferation until day 4 (d4), or to inhibit the allogeneic mixed lymphocyte reaction (MLR) of the same pPBMC batch until day 7 (d7). Representative CD3+ T-cell proliferation kinetics (Modfit analysis) in the absence (top histograms), or presence of regulatory MSPCs (bottom histograms) indicating maximum number of proliferated populations (top histograms) and the effect of MSPC-mediated inhibition of T-cell proliferation (bottom histograms) are shown